EP3885344B1

EP3885344B1 - Pyrimidine compounds, preparation method therefor and pharmaceutical uses thereof

Info

Publication number: EP3885344B1
Application number: EP21166266.3A
Authority: EP
Inventors: Yueheng Jiang
Original assignee: Inventisbio Co Ltd
Current assignee: Inventisbio Co Ltd
Priority date: 2014-11-05
Filing date: 2015-11-05
Publication date: 2025-03-26
Anticipated expiration: 2035-11-05
Also published as: CA2966376A1; JP2017533266A; IL286165B2; EP3216786A4; US10179784B2; CN115403565B; US20240208967A1; IL252036A0; CN107548391A; KR102560052B1; CN111170998B; CA2966376C; JP6637058B2; EP3216786A1; EP3885344A2; CN111170998A; CN105085489B; CN115403565A; KR20170101908A; CN105085489A

Description

FIELD OF THE INVENTION

The present invention pertains to the field of pharmaceutical chemistry, and particularly, to pyrimidine compounds and pharmaceutically acceptable salts thereof and a pharmaceutical composition containing the same and pharmaceutical use of the same. In particular, the present invention relates to pyrimidine compounds and pharmaceutically acceptable salts thereof and a pharmaceutical composition containing the compounds or pharmaceutically acceptable salts thereof and a pharmaceutical composition comprising the compounds and pharmaceutically acceptable salts thereof for use in treating diseases mediated by EGFR in the form of activated mutants and/or resistant mutants.

BACKGROUND

Cancer is becoming the deadliest "killer" to human beings. In recent years, the total number of people died for cancer is close to 2 million each year in China. Although a variety of discovery of treatment pathways and drugs have brought hope for cancer patients, these conventional treatments still have many drawbacks, such as large side effect, poor treatment effect, tumor recurrence, metastasis and so on. There is an urgent need for new treatment techniques to improve the low success rate of cancer treatment. The recent emergence of individualized chemotherapy and targeted therapy has brought new hope to lung cancer treatment. Tumor molecular targeted therapy is a treatment method in which the key molecules that closely relate to the tumor growth will selectively kill the tumor cells through chemical or biological means. Targeted therapy has many characteristics, such as high specificity, high selectivity and mild side effects. When targeted therapy is used in combination with traditional chemotherapy, radiotherapy or tumor immunization, the efficacy can be greatly enhanced and the postoperative recurrence can be reduced. Tumor targeted therapy has rapidly develop in recent years, and becomes the emerging field of cancer treatment and future development trend.
Protein tyrosine kinases (PTKs) are a class of protein enzymes that can catalyze the phenolic hydroxyl phosphorylation on tyrosine residue of a variety of important proteins, thereby activating the biological activity of functional proteins. This reaction process plays a very important role in the intracellular signal transduction pathway, for it regulates a series of physiological and chemical processes such as cell growth, differentiation and death. Protein tyrosine kinase dysfunction can cause a series of diseases in the body. There are many studies showing that the activation of more than half of the original cancer gene and oncogene are associated with protein tyrosine kinase, and protein tyrosine kinase abnormal expression can lead to disorders of cell proliferation regulation, thereby leading to tumor genesis. In addition, tyrosine kinase abnormal expression is also closely associated with tumor invasion and metastasis, tumor neovascularization, tumor resistance to chemotherapy. Tyrosine kinase has become a very important target for the development of antitumor drugs.
Epidermal growth factor receptor (EGFR) is a receptor tyrosine protein kinase, and a transmembrane protein in the ErbB receptor family.
EGFR regulates proliferation, survival, adhesion, migration and differentiation of cells, which is hyperactivated or sustained in a variety of tumor cells, such as lung cancer cells, breast cancer cells, prostate cancer cells. Abnormal activation of EGFR plays a key role in tumor transformation and growth. Blocking activation of EGFR has been clinically proven as one of the effective targeted therapy methods for tumor cell. EGFR was expressed in 50% of NSCLC (non-small cell lung cancer) cases, which makes EGFR and family members thereof a major candidate for targeted therapy. Gefitinib and erlotinib are the first generation of small molecule inhibitors of EGFR, and primarily used as drugs for treating advanced NSCLC. Clinical results show that gefitinib or erlotinib has effect on about 10% of white NSCLC and about 35% of Asian NSCLC patients. The analysis shows that the response rate to EGFR-tyrosine kinase inhibitor (TKI) in most NSCLC patients with EGFR activation mutations was significantly higher than that in EGFR wild type of NSCLC patients.
However, clinical studies have shown that many patients soon (12-14 months) have been resistant to these small molecule inhibitors of EGFR, i.e., acquired drug resistance. Gatekeeper residue of T790M mutation is a mutation point in EGFR 20 exon and is one of the major mechanisms leading to drug resistance. Studies on a new generation of inhibitor for these EGFR mutations have recently been very successful. Afatinib is a potent and irreversible double inhibitor of EGFR and human epidermal growth factor receptor 2 (HER2) tyrosine kinases. Other similar multi-target, highly active and irreversible inhibitors, such as canertinib, and dacomitinib are also in later clinical trials. These novel second-generation irreversible inhibitors have a strong inhibitory effect on EGFR with L858R and T790M mutants, and have a significant effect on gefitinib or erlotinib-resistant cancer patients. However, these second-generation EGFR mutant inhibitors also have a strong inhibitory effect on wild-type EGFR (WT-EGFR). Clinical studies have shown that the inhibition of wild-type EGFR can lead to drug toxicity and side effects in most patients, such as rash or diarrhea in the human body.
In order to overcome the toxicity and side effects of the second-generation EGFR inhibitors, it is necessary to reduce the inhibitory effect on wild-type EGFR (WT-EGFR). A new generation (i.e. the third generation) of EGFR inhibitors should remain a strong inhibition against EGFR L858R activated mutants, Exon19 deletion activated mutants and T790M resistant mutants, and show a relatively low inhibitory effect on WT-EGFR and other tyrosine protein kinase receptors. Such compounds can be used not only in the treatment of cancer patients with a resistance to EGFR L858R-activated mutants and Exon19 deletion-activated mutants, but also in the treatment of cancer patients with EGFR-T790M resistant mutants resulting to the resistance against the first-generation EGFR inhibitors such as gefitinib, erlotinib or icotinib. The third-generation EGFR inhibitor, AZD9291, has a beneficial clinical effect, but its major metabolite, AZ5104, has a strong inhibitory effect on wild-type EGFR (WT-EGFR), which is the most probable incentive inducing the most common side effects such as a clinically common rash and diarrhea.
The present invention shows pyrimidine compounds that have a high inhibitory activity against EGFR mutant(s), but only relatively low inhibitory effects on wild-type EGFR. The compounds of the present invention have good physicochemical properties and safety toxicity parameters. Such compounds will have a better effect in the treatment of cancer with EGFR-activated mutants and/or EGFR-resistant mutations.
The present invention relates to certain pyrimidine compounds and pharmaceutically acceptable salt thereof, and can be used for the treatment or prevention of a disease or condition mediated by some mutated forms of epidermal growth factor receptors (e.g., L858R activated mutants, Exon19 deletion activated mutants, and T790M resistant mutants). Such compounds and pharmaceutically acceptable salts thereof can be used for the treatment or prevention of many different cancers. The present invention also relates to a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof; and said compound and pharmaceutical composition for use in treating diseases mediated by EGFR in the form of activated and/or resistant mutants.
In the following list of documents, the patent or non-patent documents (journals, magazines, manuals and books, etc.) that are closest to patent applications are cited:

1. New England Journal of medicine, 2008, vol. 358, pp. 1160-1174;
2. Chemical and Biophysical Research Communications, 2004, vol. 319, pp. 1-11;
3. Science, 2004, vol. 304, pp. 1497-1500;
4. New England Journal of medicine, 2004, vol. 350, pp. 2129-2139;
5. Molecular Cancer Therapeutics, 2014, vol. 13, pp. 1468-1479;
6. Journal of Medicinal Chemistry, 2014, vol. 57, pp. 8249-8267;
7. WO2013014448A1 , corresponding to CN103702990A ;
8. WO2013108754Al ;
9. CN103374000A ;
10. CN103804303A ;
11. WO2013184766A1 ; and
12. WO2009051822A1 .

It should be stated that the above-mentioned patent and non-patent documents are only representative documents and are not a complete list of all the relevant literature.
The current EGFR-TKI does still not solve the clinical problems caused by drug resistance, and the most of existing drugs are EGFR reversible or irreversible inhibitors based on quinazoline or quinolinamine as the basic nucleus, and they are still inevitably brought to the side effects of poor selectivity to EGFR wild-type cells. Therefore, there is an urgent need for a new type of compounds, especially compounds with novel skeletons, so as to solve problems such as poor drug resistance and selectivity.

SUMMARY OF THE INVENTION

In a first aspect the present invention provides a pyrimidine compound represented by:
or a pharmaceutically acceptable salt thereof. The compounds can inhibit the variants of epidermal growth factor receptor (EGFR) protein kinases, and therefore can inhibit the growth of a variety of tumor cells effectively. The compounds can be used to prepare antitumor drugs, used for the treatment or prevention of various different cancers. The compounds can overcome the drug resistance induced by the existing gefitinib, erlotinib and so on. More particularly, the compounds can be used to prepare drugs for treating or preventing diseases, disturbances, disorders or conditions mediated by EGFR variants (such as L858R activated mutants, Exon19 deletion activated mutants and/or T790M resistant mutants).
In a further aspect, the present invention provides a pharmaceutical composition comprising the above pyrimidine compound and/or a pharmaceutically acceptable salt thereof, and one or more pharmaceutical excipients.
In a further aspect, the present invention provides said pyrimidine compound and/or pharmaceutically acceptable salt thereof, and the above pharmaceutical composition for use in treating or preventing diseases, disturbances, disorders or conditions mediated by a variant EGFR, particularly one or more cancers.
The compound or pharmaceutical composition for use of the present invention may be used in a combined treatment of cancer, that is to say, a method for treating cancer by using one or more of the above pyrimidine compound and pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to the present invention in combination with conventional surgery, radiotherapy, chemotherapy or tumor immunotherapy.
The pharmaceutical composition comprises a therapeutically effective amount of one or more of the compound of the present invention and/or a pharmaceutically acceptable salt thereof, and one or more pharmaceutical excipients. The pharmaceutical composition is a medicament for the treatment or prevention of diseases, disturbances, disorders or conditions mediated by the EGFR in the form of activated mutant or resistant mutant, in particular, for the treatment or prevention of one or more cancers.
The above-mentioned medicaments, according to the objective of the treatment, may be in a variety of pharmaceutical forms, generally including: tablets, pills, capsules, granules, suspensions, solutions, creams, ointments, powders, suppositories, aerosols, injections etc.
The use in treating or preventing a disorder or disease mediated by EGFR in the form of activated mutant or resistant mutant includes that the disorder or disease is ovarian cancer, cervical cancer, colorectal cancer (e.g., colon adenocarcinoma), breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer (e.g., non-small cell lung cancer), hepatocellular carcinoma, gastrointestinal stromal tumors (GIST), thyroid cancer, cholangiocarcinoma, endometrial cancer, renal carcinoma, anaplastic large cell lymphoma, acute myeloid leukemia (AML), multiple myeloma or mesothelioma.
In the present invention, the EGFR in the form of activated mutant or resistant mutant may be, for example, a L858R activated mutant, an Exon19 deletion activated mutant and/or a T790M resistant mutant. Thus, the disease, disturbance, disorder or condition mediated by EGFR in the form of activated mutant or resistant mutant can be, for example, a disease, disturbance, disorder or condition mediated by L858R activated mutant, Exon19 deletion activated mutant and/or T790M resistant mutant.
The compounds according to the invention, or pharmaceutical composition according to the invention can be particularly used for treating or preventing disease, disturbance, disorder or condition mediated by EGFR in the form of activated mutant or resistant mutant, such as a disease, disturbance, disorder or condition mediated by L858R activated mutant, Exon19 deletion activated mutant and/or T790M resistant mutant, and may be used, for example, for treating a cancer patient who has been resistant to gefitinib, erlotinib, or icotinib.
A combination therapy for treating cancer, comprising administering to a subject in need of treatment a therapeutically effective amount of the compound of the present invention or pharmaceutically acceptable salts thereof, or a therapeutically effective amount of a pharmaceutical composition according to the invention, in combination with conventional surgical therapy, radiotherapy, chemotherapy or antitumor immunotherapy is also possible.
The compounds according to the present invention may be administrated in parallel, concurrently, sequentially, or separately with the chemotherapy or antitumor immunotherapy. The chemotherapy or immunotherapy includes one or more of the following types of antitumor agents: alkylating agent (e.g., carboplatin, oxaliplatin, cisplatin, cyclophosphamide, nitrosourea, nitrogen mustard, melphalan), antimetabolite (e.g. gemcitabine), and anti-folic acid agent (e.g., 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytarabine, hydroxyurea), topoisomerase inhibitor (e.g., etoposide, topotecan, camptothecin), anti-mitotic agent (e.g., vincristine, vinblastine, vinorelbine, paclitaxel, taxotere), anti-tumor antibiotic (e.g., doxorubicin, bleomycin, doxorubicin, daunomycin, mitomycin C, actinomycin), antiestrogen drug (e.g., tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene), anti-androgen drug (e.g., bicalutamide, flutamide, nilutamide), LHRH antagonist or LHRH agonist (e.g., goserelin, leuprolide, and buserelin), aromatase inhibitor (e.g., anastrozole, letrozole), CYP17 cleavage enzyme inhibitor (such as abiraterone), anti erbB2 antibody trastuzumab [Herceptin], anti-EGFR antibody cetuximab [Erbitux]; inhibitor of tyrosine kinase, serine/threonine kinases (e.g., imatinib, nilotinib, sorafenib, trametinib, crizotinib); cyclin-dependent kinase inhibitor (e.g., CDK4 inhibitor, palbociclib), anti-human vascular endothelial growth factor antibody of bevacizumab (Avastin) and VEGF receptor tyrosine kinase inhibitor (apatinib); antitumor immunotherapy, such as anti-PD-1 antibody (pembrolizumab, nivolumab), anti-PD-L1 antibody, anti-LAG-3 antibody, anti-CTLA-4 antibody, anti-4-1BB antibody, anti-GITR antibody, anti-ICOS antibody, interleukin 2.

ADVANTAGEOUS EFFECTS

The pyrimidine compound of the present invention shows a high inhibitory activity against one or more of EGFR-activated mutant or resistant mutant and a relatively low inhibition against a wild-type EGFR. The compound of the present invention has a good physicochemical property and safety/toxicity parameter. Such compounds have a better clinical effect in the treatment of disease (including cancer) mediated by EGFR-activated mutant and/or drug-resistant mutant. Compared with AZD9291, such compounds have no or only a relatively low level of AZ5104 (a demethylated metabolite of AZD-9291) in animal in vivo experiments.

EXAMPLES

The following examples further illustrate the invention, but these examples are not to limit the scope of the invention.

Example 6 (not part of the claimed invention)

1. Synthesis of intermediate 006-2

The intermediate 001-5 (1.3 g, 8.73 mmol), 13 mL of DME, FeCl₃ (1.414 g, 8.72 mmol) and the intermediate 006-1 (974 mg, 7.43 mmol) were added sequentially to a 100 mL three-necked flask under nitrogen atmosphere, and the reaction mixture was in an oil bath at 64°C overnight. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered. The filter cake was washed three times with 20 mL of methanol and the organic phases were combined, concentrated to dryness and 1.0 g of intermediate 006-2 (47%) was obtained as a yellow solid. LCMS: 244.1.

2. Synthesis of intermediate 006-4

The intermediate 006-3 (100 g, 708.5 mmol) and 800 mL concentrated sulfuric acid (H₂SO₄) were sequentially added to a 2000 mL three-necked flask under nitrogen atmosphere, and the reaction mixture was cooled to 0°C. Potassium nitrate (KNO₃) (71.6 g, 708.2 mmol) was added in batches at 0-10°C for 1 h and then the reaction was maintained at room temperature for overnight. After the reaction was complete, 2000 mL of ice water was added to quench the reaction. The reaction mixture was adjusted to pH 10 with aqueous ammonia at low temperature and extracted three times with 1 L of dichloromethane (DCM). Then, the organic phases were combined, washed three times with 3 L saturated brine, dried over anhydrous sodium sulfate and then subjected to rotary evaporation. The crude product was purified by silica gel column chromatography (eluent, ethyl acetate (EA): petroleum ether (PE) = 1:4 - 1: 1) and eluent was concentrated to give 79 g of the intermediate 006-4 (60%) as a yellow solid. LCMS: 187.0.

3. Synthesis of intermediate 006-5

The intermediates 006-2 (75 mg, 307.8 mmol) and 006-4 (57.4 g, 308.4 mmol), 975 mL of isopropyl alcohol, and p-toluenesulfonic acid (63.7 g, 369.9 mmol) were sequentially added into a 2 L four-necked flask under nitrogen atmosphere, and the reaction was heated and maintained at 105°C for 5 h. The reaction mixture was cooled to room temperature and filtered, and the filter cake was washed with 750 mL of isopropanol three times. The filter cake was washed three times with 750 mL of acetonitrile and dried to give 75 g of the intermediate006-5 (62%) as a yellow solid. LC-MS: 394.1.

4. Synthesis of intermediate 006-7

The intermediates 006-5 (500 mg, 1.27 mmol) and 006-6 (147 mg, 1.65 mmol) and K₂CO₃ (526 mg, 3.81 mmol) were added into a 50 mL single-necked flask, and NMP (20 mL) was added thereto at room temperature. Under nitrogen protection, the oil bath was heated to 100°C. After 2 h of reaction, the mixture was cooled to room temperature. The reaction solution was dropped into 100 mL of a mixture of ice and water, and filtered by suction. The filter cake was collected, was washed three times with 50 mL water and dried to give 430 mg of the intermediate 006-7 (68%) as a red solid. LC-MS: 463.2.

5. Synthesis of intermediate 006-8

DCM : MeOH = 1:1 (20 mL) was added to a 250 mL single-necked flask at room temperature, followed by addition of the intermediate 006-7 (400 mg, 0.86 mmol), ammonium formate (400 mg, 6.34 mmol) and palladium on carbon containing water (400 mg, 5% Pd). The reaction was carried out at room temperature for 3 h. The reaction mixture was filtered, and the filtrate was collected and subjected to rotary evaporation to give a crude product which was purified by silica gel column chromatography (eluent: DCM) to give 350 mg of the intermediate 006-8 (94%) as a pale red solid. LC-MS: 433.2.

6. Synthesis of final product 6

Anhydrous ethanol (20 mL) was added into a 100 mL three-necked flask at room temperature, and then the intermediate 006-8 (340 mg, 0.787 mmol) and DIPEA(203 mg, 1.57 mmol) were added. The reaction mixture was cooled to 0°C in an ice-water bath followed by dropping acryloyl chloride (70 mg, 0.787 mmol). The reaction was carried out at 0°C for 2 h, and then was quenched by adding 2 mL of water. The reaction mixture was subjected to rotary evaporation to give a crude product which was purified by silica gel column chromatography (eluent: DCM : MeOH = 30 : 1). Eluent was concentrated to give compound 6.
The product 6 was dissolved in 4 mL of acetonitrile. Excess of concentrated hydrochloric acid was added dropwisely and the resulting mixture was concentrate directly. The crude was subjected to freeze drying to give 26.3 mg of hydrochloride of the product 6 (6%) as a yellow solid. LCMS (parent molecule) C₂₇H₃₀N₆O₃ (ES, m/z): [M+H]⁺ = 487. ¹H-NMR (300MHz, D₂O, ppm) δ 3.13 (s, 3 H), 3.21 (s, 3 H), 3.32-378 (m, 7 H), 3.89 (s, 3 H), 5.87-5.90 (d, J = 11.4 Hz, 1 H), 6.32-6.41 (m, 2 H), 6.74-6.77 (d, J = 6.0 Hz, 1 H), 6.91-6.94 (m, 1 H), 7.13-7.27 (m, 3 H), 7.57-7.65 (m, 2 H), 7.90 (s, 1 H), 7.99 (s, 1 H).

Example 8 (not part of the claimed invention)

1. Synthesis of intermediate 008-2

The intermediates006-5 (1.0 g, 2.54 mmol) and 008-1 (0.430 mg, 3.31 mmol) and K₂CO₃ (1.05 g, 7.63 mmol) were sequentially added into a 50 mL single-necked flask, and NMP (20 mL) was added thereto at room temperature. Under nitrogen protection, it was heated to 100°C in oil bath. After 2 h of reaction, the mixture was cooled to room temperature. The reaction solution was dropped into 100 mL of a mixture of ice and water, and filtered by suction. The filter cake was collected, washed three times with 50 mL water and dried to give 0.8 g of the crude product 008-2 as a red solid.

2. Synthesis of intermediate 008-3

DCM/MeOH (1:1, 20 mL) was added to a 250 mL single-necked flask at room temperature, followed by the addition of the intermediate 008-2 (800 mg, 2.38 mmol), ammonium formate (800 mg, 12.7 mmol) and palladium on carbon containing water (0.800 g, 5% Pd). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was filtered, and the filtrate was collected and subjected to rotary evaporation to give a crude product which was purified by silica gel column chromatography (eluent: DCM/MeOH = 30: 1). Eluents were combined and concentrated to give 0.650 g of the intermediate 008-3 (86%) as a pale red solid.

3. Synthesis of compound 8

Anhydrous ethanol (20 mL) was added into a 100 mL three-necked flask at room temperature, and then the intermediate 008-3 (300 mg, 0.63 mmol) and DIPEA (163 mg, 1.27 mmol) were added thereto. The reaction mixture was cooled to 0°C in an ice-water bath followed by addition of a solution of acryloyl chloride (56 mg, 0.6 mmol) in 2 mL of anhydrous THF. The reaction was stirred at 0°C for 1 h, and then was quenched by adding 2 mL of water. The reaction mixture was subjected to rotary evaporation to give a crude product which was purified by silica gel column chromatography (eluent: DCM/ MeOH = 30: 1). Eluents were combined and concentrated to give compound 8.
The compound 8 was dissolved in 4 mL of anhydrous acetonitrile, and a solution of methanesulfonic acid (65.6 mg, 6.8 mmol) was added dropwisely. After the reaction was carried out for 2 h at room temperature, a yellow solid was precipitated and the mixture was filtered by suction. The solid cake was collected and dried to give 53 mg of sulfonate of the compound 8 (7%) as a yellow solid. LCMS (parent molecule) C₃₀H₃₇N₇O₂ (ES, m/z): [M+H]⁺= 528. ¹H-NMR (300 MHz, DMSO-D₆, ppm) δ 9.85-9.86 (m, 1 H), 9.55-9.64 (m, 1 H), 8.73 (s, 1 H), 8.26 (s, 3 H), 7.57-7.60 (d, J = 8.1 Hz, 1 H), 7.37-7.39 (m, 2 H), 7.29-7.28 (m, 1 H), 7.05 (s, 1 H), 6.82-6.95 (m, 1 H), 6.29 (s, 1 H), 5.75-5.79 (d, J = 12.3 Hz, 1 H), 3.92 (s, 3 H), 3.89 (s, 3 H), 3.27 (m, 4 H), 3.15 (m, 4 H), 2.69 (s, 3 H), 2.31 (s, 3 H), 1.17-1.22 (m, 6 H).

Example 22 (not part of the claimed invention)

1. Synthesis of intermediate 022-3

The intermediate 022-1 (10 g, 42.1 mmol) as raw material was dissolved in 100 mL of dichloromethane in a 250 mL three-necked flask at room temperature under a nitrogen atmosphere, and cyclobutylamine hydrochloride (4.7 g, 50.2 mmol) was added thereto. Then, the reaction was carried out for 1h at room temperature, followed by that the reaction mixture was cooled to 0°C. Sodium triacetoxyborohydride (13.4 g, 63.2 mmol) was added to the reaction system in batches, and the reaction temperature was raised to room temperature and the reaction was stirred overnight. The reaction system was adjusted to pH 8-9 with anhydrous sodium carbonate aqueous solution the next day, and the mixture was extracted three times with 200 mL of methylene chloride. The organic phases were combined and washed once with 300 mL of saturated brine. The organic phases were dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (eluent: EA / PE = 1: 5) to give 4.7 g of intermediate 022-3 as a yellow oil. LCMS: 279.2.

2. Synthesis of intermediate 022-4

The intermediate 022-3 (3 g, 10.8 mmol) as raw material was dissolved in 30 mL of anhydrous methanol in a 100 mL single-necked flask at room temperature. Palladium on carbon containing water (3 g, 5% Pd) was added and the reaction system was exchanged with hydrogen 3 times. The reaction was carried out overnight at room temperature under hydrogen. After the reaction was completed, the mixture was filtered by suction and the filtrate was collected and concentrated to give 1.2 g of crude product 022-4 as yellow oil. LCMS: 113.1.

3. Synthesis of intermediate 022-6

The reaction steps and conditions for synthesizing compound 022-6 were completely the same as those from the first step to the second step in Example 8, except that the intermediate 008-1 as a raw material in the first step of the example 8 was replaced with the intermediate 022-4 in the first step. LCMS: 445.3.

4. Synthesis of compound 22

The intermediate 022-6 (250 mg, 0.55 mmol) as raw material was dissolved in 50 mL of anhydrous THF in a 100 mL single-necked flask at room temperature under a nitrogen atmosphere, and N,N-diisopropylethylamine (DIPEA) (141.8 mg, 1.10 mmol) was added thereto. After the reaction mixture was cool to 0 °C, acryloyl chloride (48.9 mg, 0.540 mmol) was added dropwisely to the reaction system at 0°C. The reaction system was heated to room temperature, and stirred for 1 h. After the reaction was completed, the reaction was quenched by the addition of 2 mL of water and the mixture was concentrated to dryness. The resulting residue was purified by prep-HPLC (column: Xbridge Prep RP18, 5 um, C18, 19 x 150 mm; mobile phase: 0.05% ammonia +10 mmol of ammonium bicarbonate) / acetonitrile; 77% acetonitrile to 81% acetonitrile, 4 min; 5 mL/min; detection wavelength: 254 nm), concentrated and freeze dried to give 9.8 mg of product 22 (4%) as a yellow solid. LCMS (parent molecule) C₂₀H₃₁N₇O;: (ES, m/z): 510 [M+H]⁺. ¹H-NMR: (DMSO-D₆, 300 MHz, ppm) δ 9.29 (s, 1 H), 8.34-8.32 (m, 2 H), 8.26-8.24 (d, J =5.4 Hz, 1 H), 7.92 (s, 1 H), 7.76 (s, 1 H), 7.51-7.49 (m, 1 H), 7.23-7.16 (m, 2 H), 7.13~7.12 (d, J=5.1 Hz, 2H), 6.55-6.46 (m, 1 H), 6.22 (s, 1 H), 6.17-6.16 (d, J =2.1 Hz, 1 H), 5.70-5.66 (m, 1 H ), 3.88 (s, 3 H), 3.84 (s, 3 H), 3.60-3.3.55 (m, 2 H), 3.41-3.34 (m, 1 H), 3.17-3.13 (t, J =6.9 Hz, 4 H), 2.00-1.95 (m, 2 H).

Example 28 (not part of the claimed invention)

1. Synthesis of intermediate 028-2

Under nitrogen protection, anhydrous N,N-dimethylformamide (DMF) (150 mL) was added to a 250 mL single-necked flask and then the intermediate 028-1 (375.5 mg, 3.05 mmol) was added thereto. The reaction mixture was cooled to 0°C under an ice-water bath. After sodium hydride (NaH) (65%, mineral oil mixture) (564 mg, 15.2 mmol) was added the reaction mixture in batches, the reaction temperature was raised to room temperature and the reaction was carried out for 0.5 h. Then, the intermediate 006-5 (1.0 g, 2.54 mmol) was added into the reaction mixture, and the reaction was carried out at room temperature overnight. After the reaction was completed, the reaction was cooled to 0°C and quenched by the adding 2 mL of MeOH and subjected to rotary evaporation. The crude product was purified by silica gel column chromatography (eluent: DCM / MeOH = 8:1 - 6:1) and subjected to rotary evaporation to give 0.8 g of the intermediate 028-2 (68%) as a yellow solid. LCMS: 461.2.

2. Synthesis of compound 28

The two reaction steps and conditions of synthesizing compound 28 were completely the same as those of the second step in Example 8 and the fourth step in Example 22 respectively. LCMSLCMS (parent molecule) C₂₇H₂₈N₆O₃: (ES, m/z): 485 [M+H ]⁺. ¹H-NMR (300 MHz, DMSO-D₆, ppm): δ 2.32 (s, 3 H), 3.07-3.11 (dd, J = 6.9 Hz, J = 13.2 Hz, 2 H), 3.76-3.80 (dd, J = 8.1 Hz, J = 14.4 Hz, 2 H), 3.84 (s, 3 H), 3.88 (s, 3 H), 4.83-4.87 (m, 2 H), 5.70-5.74 (d, J = 12 Hz, 1 H), 6.20-6.26 (d, J = 16.8 Hz, 1 H), 6.56 (s, 1 H), 6.65-6.76 (m, 1 H), 7.12-7.26 (m, 2 H), 7.49-7.52 (d, J = 8.4 Hz, 1 H), 7.88 (s, 1 H), 8.26-8.30 (m, 2 H), 8.46 (s, 1 H), 8.67 (s, 1H), 9.28 (s, 1 H).

Example 29

1. Synthesis of intermediate 029-2

The reaction step and condition of synthesizing compound 029-2 were completely the same as those the first step in Example 28, except that anhydrous DMF in the first step of Example 28 as a solvent was replaced with anhydrous tetrahydrofuran in this step. LCMS: 463.2.

2. Synthesis of compound 29

The reaction steps and conditions of synthesizing compound 29 and the methanesulfonate (MsOH)₃ of compound 29 were the same as those of the second step and the third step in Example 8. LCMSLCMS (parent molecule) C₂₇H₃₀N₆O₃: (ES, m/z): [M+H]⁺ = 487. ¹H-NMR (300MHz, DMSO-D₆, ppm) δ 2.28-2.36 (m, 6 H), 2.95-3.05 (m, 6 H), 3.65-3.73 (m, 2 H), 3.86 (s, 3 H), 3.93 (s, 3 H), 4.51 (s, 2 H), 5.75-5.78 (d, J = 10.1 Hz, 1 H), 6.21-6.27 (d, J=16.8 Hz, 1 H), 6.62-6.71 (m, 1 H), 7.01 (s, 1 H), 7.19-7.22 (m, 1 H), 7.28-7.33 (t, J = 7.8 Hz, 1 H), 7.37-7.39 (d, J = 6.3 Hz, 1 H), 7.58-7.61 (d, J = 8.4 Hz, 1 H), 8.17-8.18 (br s, 1 H), 8.33-8.76 (m, 2 H), 9.41 (s, 1 H), 9.61 (br s, 1 H).

Experimental Example 1

Experiment of cell growth inhibition

The compounds that were preferentially targeted for EGFR targeting certain mutations and relatively weak in wild-type EGFR were identified by determining the growth of cells. The NCI-H1975 cell line is a human non-small cell lung cancer cell containing T790M and L858R EGFR mutations, and the cell is grown in RPMI-1640 medium (GIBCO) containing 10% fetal bovine serum (FBS). The LoVo cell line is a wild-type EGFR human colon adenocarcinoma cell, and is grown in F-12K medium (GIBCO) containing 10% FBS. NCI-H2073 cell line is a wild-type EGFR human non-small cell lung cancer cell and grown in ACL-4 medium containing 10% FBS. The growth rate of NCI-H1975, LoVo and NCI-H2073 cells was detected by Cell Titer-Glo luminescence activity assay (Promega # G7572).
Briefly, trypsin was used for digesting cells in the logarithmic growth phase. 96-well plates were seeded with 50,000 LoVo or NCI-H2073 cells, 2500-3000 NCI-H1975 cells per well and provided with blank control wells containing only nutrient solution without inoculated cell, and the plates were incubated in a humidified incubator with 5% CO₂ at 37°C. After 24 hours, the DMSO solution of the different compounds was diluted with a cell culture medium at 3.16 times per time to eight different concentrations from high to low levels. The concentration of test drug in NCI-H1975 cells was from 0.03 nM to 100 nM, and that in LoVo and NCI-H2073 cells was from 3 nM to 10 µM. The cell culture medium containing the different compounds was then added to a 96-well cell plate in which one cell control well comprising cell culture medium only containing DMSO was provided. After a drug treatment for 72 hours, the cell plates were removed from the incubator and allowed to stand at room temperature for 30 minutes. Next, Cell Titer-Glo reagent was added to the wells and the 96-well cell plate was shaken at room temperature for 10 minutes to induce cell lysis. The 96-well cell plate was placed on the bench for 2 minutes to stabilize the luminescence signal. Finally, the 96-well cell plate was placed in an EnVision Multi-labeled Microplate Reader (PerkinElmer), and the signal was read with an integral time of 0.5 seconds.
Formula: Percentage of cell growth inhibition% = (maximum signal - compound signal) / (maximum signal - minimum signal) * 100%
The maximal signal was obtained from the cell control well which were treated with the DMSO solution having no any compound;
The compound signal was obtained from the drug-treated cell wells to which the compound was added;
The minimum signal was obtained from a blank control well to which no cells and only nutrient solution was added.
The cell growth inhibition curve was calculated by GraphPad Prism V5.0 software and the compound concentration required to give a 50% inhibition was calculated based on this data, i.e., IC₅₀ of compounds.
The results are listed in Table 1 below. Table 1: Results of compound activity

Compound # or its salt # NCI-H1975 IC₅₀ (nM) LoVo IC₅₀ (nM) NCI-H2073 IC₅₀ (nM)

29. (MsOH)₂ 13.0 2519 2520

Claims

A pyrimidine compound represented by:
or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition, comprising the compound and/or pharmaceutically acceptable salt thereof according to claim 1, and one or more pharmaceutical excipients.
The compound and/or pharmaceutically acceptable salt thereof according to claim 1, for use in treating or preventing a disorder or disease mediated by EGFR in the form of an activated or resistant mutant, wherein the disorder or disease mediated by the EGFR in the form of an activated or resistant mutant is ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, cholangiocarcinoma, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple myeloma or mesothelioma.
The compound for use according to claim 3, wherein the disease or disorder is non-small cell lung cancer.
The pharmaceutical composition according to claim 2, for use in treating or preventing a disorder or disease mediated by EGFR in the form of an activated or resistant mutant, wherein the disorder or disease mediated by the EGFR in the form of an activated or resistant mutant is ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, cholangiocarcinoma, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple myeloma or mesothelioma.
The pharmaceutical composition for use according to claim 5, wherein the disease or disorder is non-small cell lung cancer.